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Human Mesenchymal Stem Cell Verification Flow Kit

Catalog # FMC020 | R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Human Bone-Marrow Derived Cells Fulfill the ISCT's Definition of Human MSCs Based on Marker Expression.
(3)
Surface Antigens Expressed by KG-1 Acute Myelogenous Leukemia Cells and Peripheral Blood Mononuclear Cells are Recognized by the Antibodies Included in the Negative Marker Cocktail.
Human MSCs Fulfill the ISCT’s definition of Human Mesenchymal Stem Cells Based on Functional Differentiation.
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Setting the standard in quality research reagents

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, mesenchymal stem cells can be analyzed for marker expression using the following procedure:

  • Incubate the cells with the provided antibodies
  • Wash the cells
  • Resuspend in Flow Cytometry Staining Buffer
  • Analyze by flow cytometry

Reagents Provided

Kit Components

Reagents supplied in the Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry Kit (Catalog # FMC020):

Positive Markers:

  • CD90-APC Mouse IgG2A; Clone Thy-1A1; 500 μL
  • CD73-CFS Mouse IgG2B; Clone 606112; 500 μL
  • CD105-PerCP Mouse IgG1; Clone 166707; 500 μL

Negative Markers:

  • Negative Marker Cocktail (500 mL) that includes:
    • CD45-PE Mouse IgG1; Clone 2D1
    • CD34-PE Mouse IgG1; Clone QBEnd10
    • CD11b-PE Mouse IgG2B; Clone 238446
    • CD79A-PE Mouse IgG1; Clone 706931
    • HLA-DR-PE Mouse IgG1; Clone L203

Isotype Controls:

  • Mouse IgG2A-APC Isotype Control; Clone 20102; 500 μL
  • Mouse IgG2B-CFS Isotype Control; Clone 133303; 500 μL
  • Mouse IgG1-PerCP Isotype Control; Clone 11711; 500 μL
  • Negative Isotype Cocktail (500 μL) that includes:
    • CD45-PE Mouse IgG1; Clone 2D1
    • CD34-PE Mouse IgG1; Clone QBEnd10
    • CD11b-PE Mouse IgG2B; Clone 238446
    • CD79A-PE Mouse IgG1; Clone 706931
    • HLA-DR-PE Mouse IgG1; Clone L203
  • 125 mL of 1X Staining Buffer

Other Supplies Required

Reagents

  • Fc receptor blocking reagents (optional)

Materials

  • Flow Cytometry/FACS™ Tubes (5 mL round-bottom polystyrene tubes)
  • Pipette Tips and Pipettes

Equipment

  • Clinical centrifuge
  • Hemocytometer
  • Flow Cytometer

Procedure Overview

  1. Wash cells with 2 mL of Staining Buffer.
wash cell with staining buffer
  1. Perform a cell count.
perform cell count
  1. Add Fc receptor blocking reagents (optional).
  2. Transfer 1 x 105 cells in 100 μL into a 5 mL Flow Cytometry tube.
transfer cells into flow cytometry tube
  1. Add 10 μL of each positive antibody and 10 μL of the Negative Marker Cocktail or add 10 μL of each positive isotype control antibody and 10 μL of the Negative Isotype Control Cocktail.
  2. Incubate the samples for 30-45 minutes at room temperature in the dark.
add positive antibody and positive isotype control antibody
  1. Wash the cells with Staining Buffer.
  2. Resuspend cell pellets in 200-400 μL of Staining Buffer.
resuspend cell pellets
  1. Analyze the cells for marker expression by flow cytometry.
Analyze the cells for marker expression by flow cytometry.

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Product Documents for Human Mesenchymal Stem Cell Verification Flow Kit

Certificate of Analysis

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